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The first structure of actin in complex with a compound that alters actin dynamics - latrunculin A - is described. The co-crystallisation of an actin-gelsolin complex with latrunculin A provides structural evidence for how latrunculin A binds in the ATP-binding cleft of actin, thus stabilising the actin monomer and preventing actin polymerisation.
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The 2 Å crystal structure of the whole Arp2/3 complex purified from bovine thymus is described. The structure of the inactive Arp2/3 complex provides a wealth of molecular detail on how the complex might act to nucleate actin filaments. It also confirmed the surprising accuracy of a large number of previous studies investigating the molecular associations between Arp2/3 subunits and how they might interact to generate new actin filaments.
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This is a cryo-electron microscopy reconstruction of the Arp2/3 complex in its activated state showing significant rearrangements compared with the inactive Arp2/3 complex. The active Arp2/3 complex reconstruction was also fitted to actin branches, showing for the first time that the Arp2 and Arp3 subunits were likely to form the first two subunits of the new daughter filament.
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Expression, purification and characterisation of the complete human Arp2/3 complex in insect cells is described. In light of recent structures, this tool is likely to be immensely powerful for future studies of the complex.
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A cryo-electron microscopy reconstruction of actin filaments bundled by fimbrin. This is the first atomic resolution model of an actin filament bundle showing the geometry and orientation of the actin-binding domains of fimbrin with respect to F-actin.
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Cryo-electron microscopy reconstructions of the utrophin actin-binding domain with F-actin are presented. The reconstructions revealed two different modes of binding: at one utrophin ABD per two actins, both CH domains contact actin subunits; however, at one utrophin ABD per actin only the first CH domain made contact with actin (see also Moores et al. (2000) [40]).
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Galkin V.E., Orlova A., VanLoock M.S., Rybakova I.N., Ervasti J.M., Egelman E.H. The utrophin actin-binding domain binds F-actin in two different modes: implications for the spectrin superfamily of proteins. J Cell Biol. 157:2002;243-251 Cryo-electron microscopy reconstructions of the utrophin actin-binding domain with F-actin are presented. The reconstructions revealed two different modes of binding: at one utrophin ABD per two actins, both CH domains contact actin subunits; however, at one utrophin ABD per actin only the first CH domain made contact with actin (see also Moores et al. (2000) [40]).
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Utrophin binds laterally along actin filaments and can couple costameric actin with sarcolemma when overexpressed in dystrophin-deficient mice
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The authors describe homology modelling of a cryo-electron microscopy reconstruction of an α-actinin dimer. A complete atomic model of α-actinin was constructed using known high-resolution structures for the α-actinin coiled-coil region, the dystrophin actin-binding domain and calmodulin EF-hands.
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2+-dependent actin binding. J Mol Biol. 310:2001;845-858 The authors describe homology modelling of a cryo-electron microscopy reconstruction of an α-actinin dimer. A complete atomic model of α-actinin was constructed using known high-resolution structures for the α-actinin coiled-coil region, the dystrophin actin-binding domain and calmodulin EF-hands.
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